Construction of a Macrophage-Tropic Subtype C HIV-1-mGreenLantern Reporter Virus for Studies on HIV-1 Replication and the Impact of Methamphetamine
HIV-1 subtype C viruses are responsible for 50% of global HIV burden. However, nearly all currently available reporter viruses widely used in HIV research are based on subtype B. We constructed and characterized a replication-competent HIV-1 subtype C reporter virus expressing mGreenLantern. mGreenL...
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| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2024-11-01
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| Series: | Viruses |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1999-4915/16/12/1859 |
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| Summary: | HIV-1 subtype C viruses are responsible for 50% of global HIV burden. However, nearly all currently available reporter viruses widely used in HIV research are based on subtype B. We constructed and characterized a replication-competent HIV-1 subtype C reporter virus expressing mGreenLantern. mGreenLantern sequences were inserted in-frame with Nef ATG in HIV-1<sub>IndieC1</sub>. As controls, we employed HIV-1<sub>IndieC1</sub>, HIV-1<sub>ADA</sub>, and HIV-1<sub>NLAD8-GFP-Nef</sub> viruses. HIV-1<sub>IndieC1-mGreenLantern</sub> (HIV-1<sub>IndieC1-mGL</sub>) exhibited characteristics of the parental HIV-1<sub>IndieC1</sub> virus, including its infectivity in TZMbl reporter cells and replication competence in macrophages. To further characterize HIV-1<sub>IndieC1-mGL</sub> virus, we tested its responsiveness to CCL2 levels, a characteristic feature of subtype B HIV-1 that is missing in subtype C. CCL2 immunodepletion inhibited the production of HIV-1<sub>ADA</sub> and HIV-1<sub>NLAD8-GFP-Nef</sub> as expected, but not that of HIV-1<sub>IndieC1-mGL</sub>, as previously reported. We also tested the effect of methamphetamine, as its effect is mediated by NF-kB and since subtype C viruses carry an additional copy of NF-kB. We found that methamphetamine increased the replication of all viruses tested in macrophages; however, its effect was much more robust for HIV-1<sub>IndieC1</sub> and HIV-1<sub>IndieC1-mGL</sub>. Our studies established that HIV-1<sub>IndieC1-mGL</sub> retains all the characteristics of the parental HIV-1<sub>IndieC1</sub> and can be a useful tool for HIV-1 subtype C investigations. |
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| ISSN: | 1999-4915 |